Reduced side-effects of adjuvant hormone therapy in breast cancer patients by complementary medicine.

UNLABELLED: A clinical investigation (representing evidence-based medicine level III) was performed to evaluate the benefit of complementary medicine in breast cancer patients undergoing adjuvant hormone therapy (HT). PATIENTS AND METHODS: The patients (n=129) were treated according to international guidelines. All patients suffered from arthralgia and mucosal dryness induced by the adjuvant HT. To reduce these side-effects, the patients were complementarily treated with a combination of sodium selenite, proteolytic plant enzymes (bromelaine and papain) and Lens culinaris lectin. On the basis of case report formulas (CRFs), self assessment of defined side-effects of HT (arthralgia and mucosal dryness) were documented before as well as 4 and 8 weeks after complementary treatment. Validation was carried out by scoring from 1 (no side-effects/optimal tolerability) to 6 (extreme side-effects/extremely bad tolerability). RESULTS: The severity of side-effects of HT was reduced by complementary treatment with sodium selenite, plant enzymes (bromelaine and papain) and Lens culinaris lectin. The mean score of symptoms declined from 4.2 (before treatment) to 3.2 (after 4 weeks of treatment) to 2.7 (after 8 weeks of treatment) for arthralgia and from 3.2 (before treatment) to 2.9 (after 4 weeks of treatment) to 2.6 (after 8 weeks of treatment) for mucosal dryness, the primary aims of this investigation. The reduction of side-effects of HT was statistically significant (p<0.001 after 4 weeks and p<0.0001 after 8 weeks). CONCLUSION: This investigation demonstrates benefits of indication-based complementary treatment in breast cancer patients, e.g. reduction of side-effects of adjuvant HT. A randomized controlled trial is planned to integrate the complementary treatment with the combination of sodium selenite, proteolytic enzymes and Lens culinaris lectin into evidence-based medicine.

[Effect of chronic alcohol abuse on the phagocytotic activity of blood monocytes]. / Einfluss chronischen Alkoholmissbrauchs auf die Phagozytoseleistung von Blutmonozyten.

The phagocytic abilities of human monocytes/macrophages have been investigated in people with chronic alcohol abuse. The blood monocytes of the patients were tested in three functional systems: Fc-receptor-mediated phagocytosis, phagocytosis via complement receptor interaction and the so-called lectinophagocytosis. In a group of 57 alcoholics, the values for the phagocytic ability were distributed over a wide range by using all three tests in comparison with a group of 23 non-drinking control persons. However, a significant change in the monocytic function by ethanol could not be observed. Also a correlation between age, liver damage or abstinent periods with respect for these forms of phagocytosis could not be evaluated. As a result of our investigations we discuss our hypothesis, that in chronic alcoholics the so-called "unspecific defense mechanisms" are not altered at all: on the contrary, sometimes they have been up-regulated. A functional defect of specific defense mechanisms, however, has been observed by other investigators. Accordingly, we assume a compensatory up-regulation of "unspecific" immune reactions in long-term alcoholics.

Importance of lectins for the prevention of bacterial infections and cancer metastases.

Adhesion of bacteria and of metastasizing tumour cells have much in common, especially the participation of lectins in this process. In the future it might be possible to inhibit the metastatic process and bacterial adhesion by blocking with lectins specific for appropriate (oligo) saccharides or glycoconjugates. Initial clinical trials are very promising.

Exercise, cancer and the immune response of monocytes.

We examined the influence of a regularly moderate endurance training program on qualitative and quantitative alterations of monocytes from 24 breast cancer patients. At the beginning, after 5 weeks and 7 months of bicycle training we analyzed the composition of leucocytes and phagocytotic activity of monocytes using two different methods under resting conditions. At the end of the study the amount of granulocytes was enhanced, but lymphocytes and monocytes were decreased. While the phagocytotic capacity of monocytes increased compared with receptor destroying enzyme-treated sheep red blood cells, phagocytosis against Anti D-loaded human erythrocytes did not change. These different results led us to suggest that physical exercise training increases the number of specific receptors in the surface membrane of monocytes.

Influence of a moderate exercise training on natural killer cytotoxicity and personality traits in cancer patients.

We have investigated the influence of exercise training on natural killer cells and psychological behaviour in cancer patients. 24 women with carcinomas of the breast performed a moderate training program two to three times a week for seven months. At the beginning, after five weeks and at the end of the study we analyzed the amount and the activity of natural killer cells (CD 56) under resting conditions. Personality traits were measured with the FPI-R questionnaire at the same time points. While the amount of natural killer cells did not change, their cytotoxic activity was increased at the end of the study. Furthermore, the discomfort decreased. After 5 weeks satisfaction of life was enhanced and at the end of the investigation we observed a strict connection of this improvement with the frequency of exercise. These results demonstrate a stimulating influence of moderate exercise training on the resting NK-cell activity even in cancer patients.

Cell electrophoretic discrimination of Staphylococcus saprophyticus strains of different origin.

Staphylococcus saprophyticus isolates (n = 10) from urinary tract infections (UTIs) could be reproducibly distinguished from S. saprophyticus strains from respiratory tract infections (n = 5) by distinct electrophoretic mobility. Thus, the method of "bacteriopheresis" may offer a new approach to discriminate pathogenic microbial strains of different origin with respect to defined surface structures (hemagglutinins, lectins), generating distinct surface charges.

Native and sialic acid masked Lewis(a) antigen reactivity in medullary thyroid carcinoma. Distinct tumour-associated and prognostic relevant antigens.

Forty-six medullary thyroid carcinomas (MTC) were subjected to a qualitative and quantitative characterization of native and sialic acid masked Lewis(a) (Le(a)) antigens. Immunohistochemical investigations included monoclonal antibodies (MABs) directed against alpha(2,3)-sialyl-Le(a), i.e. CA19-9 (MAB 19-9), native Le(a) (MAB anti Le(a)) and alpha(2,3)sialyl type 1 structure, i.e. CA 50 (MAB C50). To detect sialic acid masked Le(a) reactivity, MAB anti-Le(a) was also applied to native and enzymatically desialylated tissue sections with and without masking of sialic acid residues by sialic acid and sequence specific lectins. Only 7 MTC (15%) displayed a weak expression of CA19-9, while 16 (33%) showed moderate positive staining for native Le(a). Twenty-seven tumours exhibited a strong staining by the N'ase MAB anti Le(a) staining sequence. The latter could most effectively be inhibited by the simultaneous masking of alpha(2,3)-and alpha-(2,6)-linked sialic acid residues due to the competitive binding of sialic acid and sequence specific lectins: Maackia amurensis agglutinin (specific alpha(2,3)-linked sialic acid) and Sambucus nigra agglutinin (specific alpha(2,6)-linked sialic acid). Thus, in MTC the major portion of sialic acid masked Le(a) antigen reactivity is different from that detected by the MAB 19-9. The antigen reactivity is probably due to Le(a) structures containing both alpha(2,3) and alpha(2,6)-linked sialic acid residues. A highly significant correlation between the expression of CA50 and that detected by the N'ase MAB anti-Le(a) staining sequence indicates that the alpha(2,3)-sialyl type 1 chain represents a common intermediate structure within the pathway of the biosynthesis of sialylated Le(a) antigens, excluding the formation of CA19-9 via the formation of the disialyl type 1 structure. This is subsequently fucosylated to the corresponding sialic acid masked Le(a). Preliminary clinicopathological studies indicate that the sialic acid masked Le(a) antigens detected by the N'ase MAB anti-Le(a) staining sequence are related to biologically aggressive MTC.

Monoclonal antibody BW494 defines a blood group Lewisa/type 1 chain-related antigen on carcinoma-associated mucins.

The murine monoclonal antibody (MAb) BW494 defines a carbohydrate epitope on a mucin-type glycoprotein which is expressed on the majority of well-differentiated adenocarcinomas of the pancreas. In this contribution we present evidence for the detailed structural requirements of the BW494 interaction with the glycan portions on (neo)glycoproteins or (neo)glycolipids as revealed by solid-phase binding assays and inhibition assays of BW494 binding to synthetic antigens or to the affinity-isolated cancer mucin CA494. The observed cross-reactivity of spacer-linked Gal beta 1-3GalNAc beta (TF-beta) and Gal beta 1-3GlcNAc beta (type 1 chain) is indicative of an epitope that comprises a terminal Gal beta 1-3HexNAc unit. The active conformation of this epitope is critically influenced by the chemical environment of the terminal disaccharide, i.e. by adjacent aglycon or spacer moieties or by substitution with further sugar residues at the reducing end. Although a strong enhancement of antibody binding is observed for the type 1 chain derived Lea antigen, MAb BW494 is distinct in its reactivity from Lea-specific antibodies.

Monoclonal antibody FW6 defines an epitope on alpha 3/4-monofucosylated polylactosaminoglycans expressed by fetal and colon carcinoma-associated mucins.

A mouse IgM monoclonal antibody FW6 was established after immunization of mice with mucins from human amniotic fluid and was characterized with regard to its binding epitope. According to a series of biochemical criteria the epitope is located on O-linked neutral carbohydrates of M(r) 700,000 and M(r) 570,000 mucins in human amniotic fluid. The epitope is presumed to contain alpha 3/4-linked fucose and terminal beta 3/4-linked galactose that are labile to the fucosidase I from almond emulsion or to the galactosidase from bovine testes, respectively. Immunoreactive fractions of glycan alditols from human amniotic fluid mucins were partially characterized by fast atom bombardment-mass spectrometry and methylation analysis to be composed of monofucosylated polylactosamine-type deca- or nonasaccharides. According to antibody competition studies, inhibition assays with defined carbohydrates and binding assays on neoglycolipids monoclonal antibody FW6 are presumed to recognize a novel epitope that is distinct from known carbohydrate markers of the Lex/Ley family associated with colonic carcinomas. The selective reactivity of this monoclonal antibody to the majority of human colonic carcinomas and its nonreactivity to normal colonic mucosa may render this antibody as a valuable tool in cancer diagnosis or cancer treatment.

Acetylsalicylic acid has no effects on various isolated immune cells in vitro.

In addition to the well known biological effects of acetylsalicylic acid (ASA), its stimulating effect on the immune system has recently been described. In the present study, therefore, the influence of ASA on isolated leucocytes was investigated in vitro. Various concentrations of ASA, representing therapeutic concentrations, had neither an inhibitory nor a stimulating influence on the cytotoxicity of CD16-positive cells. The phytohaemagglutinin-induced proliferation of lymphocytes and the phagocytosis activity of monocytes and neutrophilic granulocytes was similarly unaffected. These results would indicate that the immunostimulating effects already described for ASA cannot be examined in isolated leucocytes in vitro and should be attributed to an interaction of various effector cells in vivo.

Monoclonal antibody SP-21 defines a sialosyl-Tn antigen expressed on carcinomas and K562 erythroleukemia cells.

Monoclonal antibody (mab) SP-21 resembles the well established mab B72.3 by high affinity binding to peptide linked sialosyl-Tn disaccharide, NeuAc alpha 2-6 GalNAc alpha, and non-reactivity to the related trisaccharide NeuAc alpha 2-6 (Gal beta 1-3)GalNAc alpha. Although mab SP-21 may be classified, accordingly, into the family of mabs defining the sialosyl-Tn antigen, its fine specificity is distinct from the reference antibody B72.3 by binding of mab SP-21 to fetal mucins from human meconium or amniotic fluid. The distinct immunoreactivity of this mab is also documented by its specific cell staining of the erythroleukemia cell line K562, where it binds to a 105 KDa glycoprotein. Moreover a subpopulation of normal lymphocytes stains to this mab SP-21.

A quantitative microassay of carbohydrate-mediated cell adhesion to glycoconjugates immobilized on polystyrene plates.

A microadhesion assay that allows the quantitative determination of carbohydrate-mediated cell adhesion to glycoconjugates immobilized on 96-well polystyrene plates has been developed. After dislodging nonadherent cells by centrifugation, specifically bound cells are quantified by colorimetric analysis of a blue formazan product generated from the dye 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide by enzymatic reduction. Carbohydrate specificity of the cell adhesion was demonstrated by inhibition analyses and the general applicability of the assay was proved with indicator cells of three different origins: mouse fibrosarcoma cells, Chang liver cells, and human breast carcinoma cells (MDA-MB 231).

Inhibition of adhesion of S-fimbriated Escherichia coli to buccal epithelial cells by human milk fat globule membrane components: a novel aspect of the protective function of mucins in the nonimmunoglobulin fraction.

We investigated the presence of factors in human milk that inhibit invasion of pathogenic bacteria. The effect of human milk fat globule membrane (HMFGM) components on adhesion of cloned S-fimbriated Escherichia coli to human buccal epithelial cells was analyzed. S fimbriae are a common feature of E. coli strains causing sepsis and meningitis in newborns and are bound to epithelia via sialyl-(alpha-2-3)galactoside structures. Human milk fat globules (HMFG) could be agglutinated by the above-mentioned bacteria. Agglutination could be inhibited by fetuin, human glycophorin, and alpha 1-acid glycoprotein. In addition, pretreatment of HMFG with Vibrio cholerae neuraminidase markedly reduced bacterium-induced agglutinations, indicating the involvement of neuraminic acid-containing glycoproteins. In contrast, lipid droplets of infant formula or artificial lipid emulsions (Intralipid) could not be agglutinated. HMFG were present in stools of breast-fed neonates as shown by indirect immunofluorescence staining with a monoclonal antibody directed against carbohydrate residues present on HMFGM. These HMFG could be agglutinated by bacteria. HMFG inhibited E. coli adhesion to buccal epithelial cells. To further characterize relevant E. coli binding structures, HMFGM components were separated by gel chromatography. The mucin fraction showed the most pronounced inhibitory effect on adhesion of S-fimbriated E. coli to human buccal epithelial cells. Our data suggest that HMFG inhibit bacterial adhesion in the entire intestine and thereby may provide protection against bacterial infection.

A novel galactose- and arabinose-specific lectin from the sponge Pellina semitubulosa: isolation, characterization and immunobiological properties.

A new lectin from the sponge Pellina semitubulosa is derived which was extracted and purified to homogeneity. The purified lectin is probably a hexamer of polypeptide chains (each M(r) 34,000) which are covalently linked via disulfide linkages; the isoelectric point is 6.1. The lectin displays the following specificities: D-galactose (50% inhibition of hemagglutination at 0.2 mM) = L-arabinose (0.2 mM) greater than D-fucose (1.5 mM) greater than D-glucose (3.0 mM). It precipitates human erythrocytes (A1, A2, A1B, B, and O) with a titer between 2(8) and 2(11) and erythrocytes from sheep and rabbits with a titer between 2(5) and 2(10). The Pellina lectin displays a strong mitogenic effect on spleen lymphocytes from mice. Immunochemical analyses revealed that both murine T- and B-lymphocytes display a capping of the lectin receptors on their cell surfaces after lectin treatment. Murine macrophages were found to endocytose the lectin. Pellina lectin at concentrations between 0.3 and 10.0 micrograms/ml potently enhances interleukin 1 (IL-1) release from mouse peritoneal macrophages and interleukin 2 (IL-2) production in mixed murine lymphocyte cultures.

The lectin from the algae Udotea petiolata: isolation, characterization and sugar binding properties.

In the present work, we describe a novel lectin which is specific for poly-N-acetyllactosamine sequences on complex N- and O-linked carbohydrate chains. This lectin was extracted and purified from the algae Udotea petiolata. The purified lectin is a monomer with a molecular mass of 65,000 and an isoelectric point of 5.6. It agglutinates normal, neuraminidase and protease-treated erythrocytes from humans irrespectively of the blood group (A, B and O) and animal erythrocytes. The Udotea lectin displays a strong mitogenic effect on human lymphocytes, especially T-cells. This lectin binds to the human serum plasma protein 8S alpha 3-glycoprotein with high affinity (ID50 0.02 microM); other species of human serum glycoproteins exhibiting a similar preponderance of complex type N-glycosylation showed also high binding capacities in the order 9.5 S alpha 1-glycoprotein greater than alpha 2-macroglobulin = beta 2 glycoprotein = immunoglobulin A greater than asialofetuin greater than alpha 1-acid glycoprotein and mucin glycopeptide (from amnion fluid). Monosaccharides and disaccharides tested do not bind to the lectin. This novel lectin will be useful for identification of N- and O-linked glycans rich in poly-N-acetyllactosamine.

Monoclonal antibody FW6 generated against a mucin-carbohydrate of human amniotic fluid recognises a colonic tumour-associated epitope.

Mucus glycoproteins of human amniotic fluid were used to generate a monoclonal antibody (MAb) FW6, which stained the intestine of a 24 week stage fetus. In adults, 76% of colonic adenocarcinomas (13/17) showed strong expression of the FW6 epitope, but it was not detected in the histologically normal mucosae adjacent to the tumours or in normal left colon mucosa. In addition, MAb FW6 stained large cell carcinomas of the lung (2/3), gastric carcinomas (5/11), and ovary adenocarcinomas (3/4). The expression in carcinomas can also be called ectopic for testing normal tissues. MAb FW6 was also reactive with pyloric mucus glands, Brunner's glands of the duodenum, Paneth cells of the ileum, pancreatic ducts, absorptive cells of the right colon, bronchiolar glands, kidney urothelia, and with a restricted number of normal mucinous tubuli of salivary gland. It was demonstrated to be under the control of the secretion gene only in intestinal Paneth cells and absorptive cells of the right colon. Comparative histochemical analysis comprising a panel of MAbs suggests that the corresponding epitope of the MAb FW6 is a type II chain related carbohydrate structure belonging to the Lex/Ley-antigen family, but is different from short chain Lex and Ley.

Invertebrate immunity: another viewpoint.

All vertebrates and invertebrates manifest self/non-self recognition. Any attempt to answer the question of adaptive significance of recognition must take into account the universality of receptor-mediated responses. These may take two forms: (1) rearranging, clonally distributed antigen-specific receptors that distinguish in the broadest sense between self and non-self, and non-self A from non-self B, latecomers on the evolutionary scene; (2) pattern recognition receptors, the earliest to evolve and still around, necessitating the requirement for induced second signals in T- and B-cell activation. Either strategy need not force upon invertebrates the organization, structure and adaptive functions of vertebrate immune systems. Thus, we can freely delve into the unique aspects of the primitive immune mechanisms of invertebrates. In contrast, using the opposite strategy which is still problematic, i.e. linking invertebrate and vertebrate defence, seems to give us an approach to universality that might eventually reveal homologous kinship.

Immunological and biological identification of tumour necrosis-like factor in sponges: endotoxin that mediates necrosis formation in xenografts.

Xenografts of the sponge Geodia cydonium in its closely related species G. rovinjensis resulted in a rapid rejection of the graft within a period of 5 days. We identified an immunoreactive tumour necrosis factor (TNF)-like activity in the xenograft (Mr of 30,000) two days after grafting. In-vivo injection of 5 micrograms human recombinant TNF-alpha induced cytotoxicity in sponge cells in the same pattern and time course as during natural xenograft rejection. Anti-TNF-alpha polyclonals were found to react with xenograft extracts, by Western blot analysis, as from day 2 after grafting. Using ELISA we detected the TNF-like activity from day 2 after grafting with peak levels at days 4 and 5, where the amount was 0.72 ng/micrograms tissue DNA. By day 1, gp27 (inhibitory aggregation factor) is already formed in the xenograft. In-vitro experiments on isolated G. cydonium cells showed that addition of purified gp27 induced the production of the TNF-like activity (up to 13.5 ng/ml). Evidence is presented that gp27 is a product of the gp180 lectin receptor. We conclude that gp27 induces TNF-like factor production, resulting in destruction and dissolution of the xenograft after 5 days.

Blood group antigen expression in medullary carcinoma of the thyroid. An immunohistochemical study on the occurrence of type 1 chain-derived antigens.

Using monoclonal antibodies (MoABs) against blood group determinants and related carbohydrate sequences, it is now possible to clarify their carcinoma-associated modulation at a molecular level. In the present study a panel of MoABs against different type 1 chain derived blood group antigens, comprising A, B, H type 1, Le(a), sialyl-Le(a) (CA 19-9), sialyl type 1 structure (CA 50), and Le(b) was used to investigate their immunoreactivity in 38 medullary carcinomas of the thyroid (MTC) and in normal thyroid tissue. The antigens were not expressed in normal follicular or C-cells but were expressed to a various extent in MTC. The studies revealed some characteristic anomalies in the frequency and patterns of tumor-associated antigen expression. The MoAB C 50 stained 32 of the 38 tumors, H type 1 (Le(d)) was demonstrated in 21 and the Le(b) antigen in 27. The Le(a)- and the A antigen were detected in 10 and 12 tumors and the B antigen in one. From the results some rules about the pathways for tumor-associated re-expression of these antigens can be deduced. Le(a) antigen expression was significantly correlated with the CA 50 and Le(b) antigens. The significant relation observed between A-, H1-, and Le(b) antigen formation in MTC suggests the existence of a carcinoma-associated fucosyltransferase committing the type 1 precursor chain along the H1-antigen pathway, and by further glycosylation to an A-, B-, or a Le(b) antigen. Comparative studies of tumor-associated H type 1 and H type 2 antigen expression revealed that H type 2 antigen synthesis was significantly related to a blood type 0 in the host. On the other hand, H1 antigen reactivity was independent of the AB0 blood type of the hosts and was also detected in H type 2 antigen-negative tumors. These findings support the proposal that even in tumor tissue, H antigen expression is still determined by the interaction of at least two different genes. Despite the occurrence of the precursor substance (CA 50) and the formation of the Le(a)- and Le(b) antigens, indicating the presence of a alpha 1,4-fucosyl-transferase (Lewis-enzyme), only two tumors showed the formation of CA 19-9. In conclusion, the investigations demonstrated the dominant re-expression of three type 1 chain-derived structures in MTC, namely H type 1, Le(b), and CA 50. These findings support the general concept demonstrated in other carcinomas, that fucosyl- and sialyltransferases are preferentially activated in MTC.(ABSTRACT TRUNCATED AT 400 WORDS)